LOCUS       sequence-298465-        5083 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046650
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(838..856)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1207..2010
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2093..3075
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3153..3182
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3204..3791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3803..3830)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3854..4045)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4046..4837,4838..4906))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4669..4688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4907..5009)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5010..5041)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgaaatttc ttgttttctt aggaatcatc acaactgtag ctgcatttca
      481 ccaagaatgt agtttacagt catgtactca acatcaacca tatgtagttg atgacccgtg
      541 tcctattcac ttctattcta aatggtatat tagagtagga gctagaaaat cagcaccttt
      601 aattgaattg tgcgtggatg aggctggttc taaatcaccc attcagtaca tcgatatcgg
      661 taattataca gtttcctgtt taccttttac aattaattgc caggaaccta aattgggtag
      721 tcttgtagtg cgttgttcgt tctatgaaga ctttttagag tatcatgacg ttcgtgttgt
      781 tttagatttc atctaaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
      841 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
      901 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
      961 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     1021 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     1081 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     1141 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     1201 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     1261 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     1321 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     1381 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     1441 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     1501 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     1561 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     1621 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     1681 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     1741 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     1801 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     1861 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     1921 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     1981 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2041 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     2101 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     2161 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     2221 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     2281 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     2341 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     2401 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     2461 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     2521 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     2581 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     2641 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     2701 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     2761 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     2821 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     2881 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     2941 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     3001 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     3061 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     3121 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     3181 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3301 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3661 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     3721 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     3781 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     3841 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3901 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3961 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4021 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4141 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4201 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4561 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     4921 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     4981 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     5041 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
